Fig. 4

RBP-Jκ KO in VSMCs blocks neointimal VSMC differentiation and prevents AVF maturation. A AVFs were created in VSMC^GFP and VSMC^{RBP−Jκ KO/GFP} mice 10 days after last dose of tamoxifen treatment (i.p., 80 mg/Kg. bw for 5 consecutive days). Representative images of double immunostaining of GFP and α-SMA were presented (A) and the GFP⁺/α-SMA⁺ cells were counted and calculated (B). C and D AVFs were created in VSMC-GFP-RBP-Jκ KO (partial KO) mice after single tamoxifen treatment (i.p., 80 mg/Kg. bw). The 1 month AVFs were immunostained with GFP/RBP-Jκ (C), GFP/α-SMA (C), and GFP/PCNA (D). E The percentage of α-SMA⁺ cells in GFP⁺ cells (representing KO of RBP-Jκ in VSMC differentiation) vs. α-SMA⁺ cells in GFP⁻ cells in the neointima (represent RBP-Jκ positive SMCs) in Panel C were calculated. F The percentage of PCNA positive cells in both GFP⁺ and GFP⁻ cells in Panel D were calculated. (n = 5; *p < 0.05; Mann–Whitney test was used for statistical analysis; white scale bars = 50 μm)